title: Fluorogenic small molecules requiring reaction with a specific protein to create a fluorescent conjugate for biological imaging--what we know and what we need to learn.
abstract: We seek fluorogenic small molecules that generate a fluorescent conjugate signal if and only if they react with a given protein-of-interest (i.e., small molecules for which noncovalent binding to the protein-of-interest is insufficient to generate fluorescence). Consequently, it is the new chemical entity afforded by the generally irreversible reaction between the small molecule and the protein-of-interest that enables the energy of an electron occupying the lowest unoccupied molecular orbital (LUMO) of the chromophore to be given off as a photon instead of being dissipated by nonradiative mechanisms in complex biological environments. This category of fluorogenic small molecules is created by starting with environmentally sensitive fluorophores that are modified by an essential functional group that efficiently quenches the fluorescence until a chemoselective reaction between that functional group and the protein-of-interest occurs, yielding the fluorescent conjugate. Fluorogenic small molecules are envisioned to be useful for a wide variety of applications, including live cell imaging without the requirement for washing steps and pulse-chase kinetic analyses of protein synthesis, trafficking, degradation, etc.
authors.affiliation: Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA, 92037; Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, 92037.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Baranczak
authors.forename: Stephen
authors.initials: S
authors.lastname: Connelly
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.forename: Sungwook
authors.initials: S
authors.lastname: Choi
authors.forename: Neil P
authors.initials: NP
authors.lastname: Grimster
authors.forename: Evan T
authors.initials: ET
authors.lastname: Powers
authors.forename: Ian A
authors.initials: IA
authors.lastname: Wilson
authors.forename: Jeffery W
authors.initials: JW
authors.lastname: Kelly
mesh_terms: D018360:Crystallography, X-Ray; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D021621:Imaging, Three-Dimensional; D008024:Ligands; D008958:Models, Molecular; D011228:Prealbumin / Q000378:metabolism*; D012997:Solvents / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013267:Stilbenes / Q000737:chemistry; D013816:Thermodynamics; D014364:Tryptophan / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D008024:Ligands; D011228:Prealbumin; D012997:Solvents; D013267:Stilbenes; D014364:Tryptophan
keywords: environmentally sensitive fluorophores; fluorescence switching; reaction regulated quenching functional groups